Bioanalytical Assay Development and Validation: Reproducible and Reliable Quantitation of Analytes

Bioanalytical assay development helps determine the design implications, limitations, favorable conditions, and suitability of a drug for the intended treatment. Before assay development and validation can be initiated, it is recommended to understand the physiochemical properties of the analyte. Bioanalytical assay development is considerably informed by the details on protein binding and in vivo/in vitro metabolism along with essential aspects of prior work on the method. Bioanalytical assays play a crucial role in the discovery and development of pharmaceutical products given their ability to discern purity, identity, performance, and potency of these compounds. Furthermore, the involved techniques allow reproducible and reliable quantitation of analytes in various biological matrices, such as plasma, urine, blood, or serum.

Assay validation is performed for the analysis of drugs, metabolites, and biomarkers to ensure that the test results are precise and accurate, and the generated data is reasonably robust in supporting drug development and other intended purposes. Several method characteristics play a significant role during each stage of assay development and validation, including specific chemistry of analyte, sample clean-up to analysis methodology, and desired method sensitivity. The selectivity and sensitivity of bioanalytical assays developed for drug and metabolite quantitation thoroughly impact the success of preclinical and clinical studies. Thus, careful sample preparation, HPLC column, and mobile phase selection, and choice of detector and quantitation method are critical during LC-MS assay development.

The purpose of assay development and validation is to find a suitable method applicable to a chosen analyte in a specific biological matrix. The analysis outcome might be inaccurate if the assay is not validated correctly and deemed unreliable. This assay inadequacy could be due to low sensitivity, selectivity, precision, accuracy or any other reason. Given the significance of this topic, the FDA has published guidance to provide guardrails around assay development and validation discussions. Generally, the assay can be used with full confidence to produce reliable data if the FDA method development guidance is followed and all its outlined criteria are met. Thus, assay development and validation are carried out with appropriate technology so that compliance with regulatory and scientific requirements can be achieved. Additionally, analytical information obtained from already existing bioanalytical assays may sometimes be effectively utilized.

Assay Development and Validation: From Drug Discovery and Preclinical to Clinical Phases

During preclinical development, bioanalytical assays assess the impact of New chemical entity (NCE) as a potential treatment on various animal species. Further assay development and validation is required to evaluate pharmacokinetic parameters, toxicity, and safety profile in humans of the drug compound that reaches the clinical stage. Sample collection, processing, and preparation should be thoroughly assessed before starting assay development to mitigate future risk factors at a later stage and validation phase.

Simply put, bioanalytical assays are imperative in improving a drug candidate’s success rate in preclinical animal studies. Robust assay development and validation during early preclinical studies is time well spent as satisfactory assays can be carried out further into development stages with minimal effort. Bioanalytical assays during clinical studies in human volunteers involve quantitation of analytes and corresponding metabolites as well as an assessment of various matrices. This analysis provides significant guidance on toxicokinetic and pharmacokinetic parameters evaluated through assay development and validation.

Bioanalytical Assay Services by NorthEast BioLab

Bioanalytical Assay Services by NorthEast BioLab

NorthEast BioLab offers comprehensive bioanalytical assay services at every stage of drug development during drug discovery, preclinical and clinical studies. Our scientists assist in assay development and validation for quantitation of analyte and metabolites for blood, plasma, serum, urine, and cerebrospinal fluid (CSF) samples among other standard matrices. Our bioanalytical method development services can be utilized for small and large molecules, such as NCE, biosimilars, peptides, antibody drug conjugates (ADC), monoclonal antibodies (mAb), and oligonucleotides.

LC-MS/MS Assay: PK, BABE, TK, Biomarkers

LC-MS-MS assays are used primarily for the clinical-grade analysis of small molecule drugs, metabolites, and biomarkers in biological fluids. NorthEast BioLab has developed and validated over 300 LC-MS/MS methods from scratch and utilized these for preclinical Tox studies and Phase I-IV clinical studies. Many of these methods were submitted to regulatory agencies as part of IND and NDA filings.

ELISA Assay: PK, BABE, TK, Immunogenicity, Biomarkers

The Enzyme-Linked Immunosorbent Assay (ELISA) helps determine and quantify macromolecules, such as proteins and peptides. Here, a capture and detection antibody pair are connected to the enzyme that accomplishes catalysis of a chemiluminescent or colorimetric substrate. Our veteran scientists at NorthEast BioLab utilize ELISA assays for determining the concentration and presence of the analyte in biological samples for a variety of drug research and development studies.

Biomarker Testing Assay: LC-MS/MS, ELISA, MSD, Luminex, qPCR

We provide comprehensive biomarker assay development and validation services, given our deep expertise in analyzing various analytes in multiplex and singleplex formats such as LC-MS/MS, ELISA, MSD ECL Assay, Luminex Assay, or qPCR for multiple therapeutic areas and species. Our biomarker testing services include method development and assay validation for novel biomarkers, development of tissue and fluid-based biomarkers, as well as GLP and GCP biomarker assessments.

Immunogenicity Testing Assay: ELISA, MSD

At NorthEast BioLab, we offer a comprehensive solution for immunogenicity testing including ADA screening, confirmation, titration, and isotyping. Our scientists diligently assess the immunogenicity of your drug product, calculate the analysis cutpoint, evaluate the formation of anti-drug antibodies, and measure the persistence of the neutralizing fractions of anti-drug antibodies.

ADME Assay

At NorthEast BioLab, we utilize in vitro and in vivo ADME studies to evaluate the dosage regimen, administration routes, and safety assessment of the drug on different types of species. A dedicated study is developed by our lab analysts to cater to the requirements of your program.

Cell-Based Proliferation Viability Assay

NorthEast BioLab performs various cell-based proliferation viability assay for metabolic activity analysis, DNA replication, antigen recognition, proliferation markers, membrane integrity, and ATP measurements. Our services in this domain include signaling and secretion, apoptosis and cell death, and cell proliferation assay.

Additional Bioanalytical Assay

At NorthEast BioLab, we offer a broad variety of cytotoxicity testing. Specifically, our scientists perform a variety of cell-based assays to determine drug Cytotoxicity and mechanism of action by understanding biochemical assay reactions triggered by the compound. An entire range of dosage of a drug compound is analyzed to understand the toxicity of a drug on animal models.

Basis of Bioanalysis

The Basics of Bioanalysis


Suitable Bioanalytical Assay Development and Validation to prove your Drug's Safety and Efficacy

At NorthEast Biolab, we recognize the pivotal significance of bioanalytical assays in drug discovery and development for measuring the efficacy and safety profile of your drug compounds in biological samples. Accordingly, our scientists develop reliable analytical methods suitable for the various stages of your drug development. For example, we offer expedited assay development and sample analysis for your drug discovery and molecule screening needs. Similarly, we perform full assay validation with strict adherence to bioanalytical method validation (BMV) regulatory guidelines during preclinical safety, toxicokinetic assessments, and clinical trials.

Additionally, we initiate our bioanalytical assay services only after developing a thorough understanding of your study requirements to yield high-quality results promptly. For small molecules, we widely utilize the LC-MS/MS method that doesn’t need extensive sample preparation. In this case, a suitable mass spectrometry setup is chosen based on the required assay sensitivity and specificity. For large molecules, we have in-depth expertise in assay validation of both customized ELISA and commercially available ELISA kits as needed. Here, the calibration range of the assay is chosen based on the expected concentration to minimize the reanalysis of study samples. Our team offers you outstanding support and maintains constant communication to smoothly meet your project-critical deadlines utilizing our robust techniques and scalable sample analysis platforms.